Enzyme promiscuity in natural environments: alkaline phosphatase in the ocean

Abhishek Srivastava, Daniel E. M. Saavedra, Blair Thomson, Juan A. L. García, Zihao Zhao, Wayne M. Patrick, Gerhard J. Herndl, Federico Baltar

Alkaline phosphatase (APase) is one of the marine enzymes used by oceanic microbes to obtain inorganic phosphorus (Pi) from dissolved organic phosphorus to overcome P-limitation. Marine APase is generally recognized to perform P-monoesterase activity. Here we integrated a biochemical characterization of a specific APase enzyme, examination of global ocean databases, and field measurements, to study the type and relevance of marine APase promiscuity. We performed an in silico mining of phoA homologs, followed by de novo synthesis and heterologous expression in E. coli of the full-length gene from Alteromonas mediterranea, resulting in a recombinant PhoA. A global analysis using the TARA Oceans, Malaspina and other metagenomic databases confirmed the predicted widespread distribution of the gene encoding the targeted PhoA in all oceanic basins throughout the water column. Kinetic assays with the purified PhoA enzyme revealed that this enzyme exhibits not only the predicted P-monoester activity, but also P-diesterase, P-triesterase and sulfatase activity as a result of a promiscuous behavior. Among all activities, P-monoester bond hydrolysis exhibited the highest catalytic activity of APase despite its lower affinity for phosphate monoesters. APase is highly efficient as a P-monoesterase at high substrate concentrations, whereas promiscuous activities of APase, like diesterase, triesterase, and sulfatase activities are more efficient at low substrate concentrations. Strong similarities were observed between the monoesterase:diesterase ratio of the purified PhoA protein in the laboratory and in natural seawater. Thus, our results reveal enzyme promiscuity of APase playing potentially an important role in the marine phosphorus cycle.

Functional and Evolutionary Ecology
External organisation(s)
University of Otago, Victoria University of Wellington, Royal Netherlands Institute for Sea Research
The ISME Journal
No. of pages
Publication date
Peer reviewed
Austrian Fields of Science 2012
106021 Marine biology
ASJC Scopus subject areas
Ecology, Evolution, Behavior and Systematics, Microbiology
Sustainable Development Goals
SDG 14 - Life Below Water
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